Novel Combinations of Natural Product Compounds for Treatment of Alzheimer Disease and Related Dementias

项目摘要/摘要 为了响应国家老龄研究所(NIA)的战略计划和PAS-19-317，我们提出了新的治疗方法 治疗阿尔茨海默病和相关痴呆(ADRDS)。我们已经开发出三种天然的新型组合 产物化合物，它调节多种类型的中枢神经系统细胞(即神经元， 小胶质细胞)与AD病理有关。这些化合物最初是通过我们以前的 NIA AMP-AD资助的AD靶点发现和药物再利用的计算系统生物学研究 我们确定了6000多个潜在目标，并对3000多个候选化合物进行了优先排序， 包括天然化合物、FDA批准的药物和小分子。这些化合物是通过 应用我们的计算方法分析AD患者死后大规模脑组织的多组学数据 系统生物学方法。在本项目中，我们建议进一步确认野生型(即C57BL6/J)的安全性 对于我们的新组合，使用已建立的AD小鼠模型(即3xTg小鼠)和治疗效果 天然产物化合物。此外，我们将验证我们新的性别-和APOE基因-的相关性- 以血清为基础的特定代谢生物标记物和额外的蛋白质生物标记物与我们的治疗效果 在这个AD小鼠模型中进行治疗。 我们假设这些天然产品组合将被证明是安全有效的治疗方法。 用于ADRD的治疗。为了解决这一假设，我们提出了以下三个目标： 目的1：评价NAD/PEITC和NADH/PEITC的全身和中枢毒性。在这个目标中，我们将首先 用LC-MS和核磁共振对化合物结构和处方稳定性进行了验证。接下来，我们将使用男性和女性 6个月龄和15个月龄的C57BL6/J，以测试我们的组合在 给药后分为急性(3天)、亚急性(7天)和慢性(28天)三个阶段。 目的2：评价这些药物组合对AD生物标志物和功能认知的治疗效果 在3xTg小鼠中的表现。在这里，我们将测量我们的治疗组合对大脑水平的影响 淀粉样β蛋白(即抗体(1-38)、抗体(1-40)、抗体(1-42))和过度磷酸化的tau。我们还将测试效果 我们的天然产品组合对神经认知能力的影响 还有莫里斯水迷宫。AIM 2中的所有研究都将在6个月大的雄性和雌性3xTg小鼠身上进行 还有15个月。 目的3：评估这些药物组合的其他基于血液的治疗生物标记物。用血 采集自15个月大的雄性和雌性3xTg小鼠的样本(目标2)，我们将验证新的基于血液的 代谢和蛋白质组学生物标记物用于评估我们联合治疗的疗效 代谢组学和蛋白质组学数据分析。 我们在体外和体内模型上的初步结果表明，在 神经保护，刺激小胶质细胞吞噬，改善3xTg小鼠的工作记忆。在此之前 这个项目的成功，我们将准备进一步证实这些组合的安全性和治疗效果 在一项临床研究中，对MCI和/或AD非常早期的化合物进行了研究。

PROJECT SUMMARY/ABSTRACT In response to National Institute on Aging's (NIA) strategic plan and PAS-19-317, we propose novel therapeuticsto treat Alzheimer disease and related dementias (ADRDs). We have developed novel combinations of three natural product compounds, which modulate complementary pathways in multiple types of CNS cells (i.e., neurons, microglia) associated with AD pathology. These compounds were originally discovered through our previous computational systems biology research on AD target discovery and drug repurposing funded by NIA AMP-AD consortium where we identified over 6000 potential targets and prioritized more than 3000 candidate compounds, including natural compounds, FDA-approved drugs, and small-molecules. These compounds were identified via analysis of multi-omics data from large-scale post-mortem brain tissues of AD patients using our computational systems biology approach. In this project, we propose to further confirm the safety in wild-type (i.e., C57BL6/J mice) and therapeutic efficacy using an established murine model of AD (i.e., 3xTg mice) for our novel combinations of natural product compounds. Additionally, we will validate the correlation of our novel sex- and APOE genotype- specific serum-based metabolic biomarkers and additional protein biomarkers with the therapeutic effects of our treatment in this AD mice model. We hypothesize that these natural product combinations will prove to be safe and effective therapeutics for treatment of ADRD. To address this hypothesis, we proposed the following three aims: Aim 1: To evaluate systemic and central toxicity of NAD+/PEITC and NADH/PEITC. In this aim, we will first validate the compound structure and formulation stability with LC-MS and NMR. Next, we will use male and female C57BL6/J aged 6 months and 15 months to test brain, liver, kidney, and heart toxicity of our combinations during the acute (3 days), subacute (7 days), and chronic (28 days) periods post-dose. Aim 2: To evaluate therapeutic effects of these drug combinations on AD Biomarkers and Functional Cognitive Performance in 3xTg mice. Here, we will measure effects of our therapeutic combinations on brain levels of amyloid beta proteins (i.e., Ab(1-38), Ab(1-40), Ab(1-42)) and hyperphosphorylated tau. We will also test effects of our combinations of natural products on neurocognitive performance using the novel object recognition test and the Morris water maze. All studies in Aim 2 will be conducted in male and female 3xTg mice aged 6 months and 15 months. Aim 3: To evaluate additional blood-based therapeutic biomarkers of these drug combinations. Using blood samples collected from 15-month-old male and female 3xTg mice (Aim 2), we will validate novel blood-based metabolic and proteomics biomarkers for evaluation of therapeutic responses of our combinations via metabolomics and proteomics data analysis. Our preliminary results on both in-vitro and in-vivo model have demonstrated great promise on significant neuroprotection, stimulation of microglial phagocytosis and improving working memory of 3xTG mice. Upon the success of this project, we will be ready to further confirm the safety and therapeutic effectsof these combinations of compounds for MCI and/or very early-stage of AD in a clinical study.